(a) Flow cytometry of splenic cells from 7- to 15-week-old control, HoipΔCd4, HoilΔCd4 and Sharpincpdm assessed for expression of TCRβ (top panels), CD4 and CD8 (middle panels), and for FOXP3, after gating on CD4+ cells (bottom panels). (b) Quantification of CD4+CD8+TCRβ+ and CD4+FOXP3+ cells in the spleen. (c) Surface expression of CD44 and CD62L on CD4+FOXP3− (top panels) and CD8+ T cells (bottom panels) in spleens of control, HoipΔCd4, HoilΔCd4 and Sharpincpdm mice. (d) Absolute numbers of CD44highCD62Llow activated cells in the CD4+FOXP3− (left graph) and CD8+ (right graph) populations from the spleens of controls, HoipΔCd4, HoilΔCd4 and Sharpincpdm mice. (e) Flow cytometry of splenic CD1d-αGalCer tetramer-positive NKT cells from 7- to 15-week-old control, HoipΔCd4, HoilΔCd4 and Sharpincpdm mice. (f) Total cell numbers and percentages of CD1d-α-galactosylceramide (α-GalCer) tetramer-positive NKT cells in the spleen. For b,d and f, each symbol represents an individual mouse; small horizontal lines indicate mean±s.d.; *P<0.05, **P<0.01, ***P<0.005 and ****P<0.001, respectively. One-way analysis of variance with a Tukey's post-hoc test for multiple comparisons was used for statistical analysis. Shpncpdm refers to Sharpincpdm mice. Data are pooled from six independent experiments with two to six mice per group (a–d) or representative of two independent experiments with three to six mice per group (e,f).